Determination of the glucocorticoids prednisone, prednisolone, dexamethasone, and cortisol in human serum using liquid chromatography coupled to tandem mass spectrometry

Glucocorticoids are an important component of immunosuppressive therapy for solid organ transplantation. A method to quantitate prednisone, prednisolone, dexamethasone and cortisol in human serum has been developed. Analysis is performed utilizing reversed-phase liquid chromatography coupled to tandem mass spectrometry. The method was validated to a lower limit of quantitation of 5.4 ng/ml for prednisone and cortisol, and 10.7 ng/ml for dexamethasone and prednisolone, with error below 7% at the lower limits. The between-day relative standard deviations ranged 2.9-7.1%. Comparison of cortisol analysis to an established method using clinical samples yielded differences below 15% for 26 of 28 determinations.     

Quantitative evaluation of the performance of different electrical tomography devices.

Two widely used electrical tomography systems, the Sheffield Mark I and the DAS-01P, were quantitatively evaluated and compared to the newly developed Goe-MF system. The performance was quantified using a hardware phantom which closely matches the real input and transfer impedances of the human thorax and allows measurements equivalent to different states of lung inflation. Our results demonstrate that adequate averaging is necessary for noise reduction for the Mark I and especially for the DAS-01P system to get meaningful results even in visualizing maximal respiratory manoeuvres. The Goe-MF system showed a notably improved signal-to-noise ratio which allows also dynamic measurements at low levels of lung volume changes, e.g., in intensive care lung injury patients.     

The ingestion of bacteria in suspension by the common carp Cyprinus carpio L.

Groups of large (65 -75 g) and small (8-17 g) common carp ( Cyprinus carpio L.) fingerlings were exposed to the bacterium Chromobacterium violaceurn in order to establish whether they could detect and ingest unattached bacteria. Small fish exposed to both bacteria and to cell-free bacterial extracts showed a significant increase in opercular beat rates, thus demonstrating that they are able to detect the presence of unattached bacteria in suspension. Examination of carp gut contents showed that the proportion of small fish ingesting bacteria increased with exposure time although no significant relationship was observed among larger fish. Significant, positive correlations between numbers of viable bacteria isolated from the intestinal tracts and concentration in the environment were observed. Possible mechanisms of bacterial ingestion are discussed.     

Functional Validation and Comparison Framework for EIT Lung Imaging

Introduction

Electrical impedance tomography (EIT) is an emerging clinical tool for monitoring ventilation distribution in mechanically ventilated patients, for which many image reconstruction algorithms have been suggested. We propose an experimental framework to assess such algorithms with respect to their ability to correctly represent well-defined physiological changes. We defined a set of clinically relevant ventilation conditions and induced them experimentally in 8 pigs by controlling three ventilator settings (tidal volume, positive end-expiratory pressure and the fraction of inspired oxygen). In this way, large and discrete shifts in global and regional lung air content were elicited.

Methods

We use the framework to compare twelve 2D EIT reconstruction algorithms, including backprojection (the original and still most frequently used algorithm), GREIT (a more recent consensus algorithm for lung imaging), truncated singular value decomposition (TSVD), several variants of the one-step Gauss-Newton approach and two iterative algorithms. We consider the effects of using a 3D finite element model, assuming non-uniform background conductivity, noise modeling, reconstructing for electrode movement, total variation (TV) reconstruction, robust error norms, smoothing priors, and using difference vs. normalized difference data.

Results and Conclusions

Our results indicate that, while variation in appearance of images reconstructed from the same data is not negligible, clinically relevant parameters do not vary considerably among the advanced algorithms. Among the analysed algorithms, several advanced algorithms perform well, while some others are significantly worse. Given its vintage and ad-hoc formulation backprojection works surprisingly well, supporting the validity of previous studies in lung EIT.     

Analysis of Morphological Parameters to Differentiate Rupture Status in Anterior Communicating Artery Aneurysms

In contrast to size, the association of morphological characteristics of intracranial aneurysms with rupture has not been established in a systematic manner. We present an analysis of the morphological variables that are associated with rupture in anterior communicating artery aneurysms to determine site-specific risk variables. One hundred and twenty-four anterior communicating artery aneurysms were treated in a single institution from 2005 to 2010, and CT angiograms (CTAs) or rotational angiography from 79 patients (42 ruptured, 37 unruptured) were analyzed. Vascular imaging was evaluated with 3D Slicer© to generate models of the aneurysms and surrounding vasculature. Morphological parameters were examined using univariate and multivariate analysis and included aneurysm volume, aspect ratio, size ratio, distance to bifurcation, aneurysm angle, vessel angle, flow angle, and parent-daughter angle. Multivariate logistic regression revealed that size ratio, flow angle, and parent-daughter angle were associated with aneurysm rupture after adjustment for age, sex, smoking history, and other clinical risk factors. Simple morphological parameters such as size ratio, flow angle, and parent-daughter angle may thus aid in the evaluation of rupture risk of anterior communicating artery aneurysms.     

Functional Regions of Interest in Electrical Impedance Tomography: A Secondary Analysis of Two Clinical Studies

Introduction

Patients with acute respiratory distress syndrome (ARDS) typically show a high degree of ventilation inhomogeneity, which is associated with morbidity and unfavorable outcomes. Electrical impedance tomography (EIT) is able to detect ventilation inhomogeneity, but it is unclear which method for defining the region of interest (ROI) should be used for this purpose. The aim of our study was to compare the functional region of interest (fROI) method to both the lung area estimation method (LAEM) and no ROI when analysing global parameters of ventilation inhomogeneity. We assumed that a good method for ROI determination would lead to a high discriminatory power for ventilation inhomogeneity, as defined by the area under the receiver operating characteristics curve (AUC), comparing patients suffering from ARDS and control patients without pulmonary pathologies.

Methods

We retrospectively analysed EIT data from 24 ARDS patients and 12 control patients without pulmonary pathology. In all patients, a standardized low-flow-pressure volume maneuver had been performed and was used for EIT image generation. We compared the AUC for global inhomogeneity (GI) index and coefficient of variation (CV) between ARDS and control patients using all EIT image pixels, the fROI method and the LAEM for ROI determination.

Results

When analysing all EIT image pixels, we found an acceptable AUC both for the GI index (AUC = 0.76; 95% confidence interval (CI) 0.58–0.94) and the CV (AUC = 0.74; 95% CI 0.55–0.92). With the fROI method, we found a deteriorating AUC with increasing threshold criteria. With the LAEM, we found the best AUC both for the GI index (AUC = 0.89; 95% CI 0.78–1.0) and the CV (AUC = 0.89; 95% CI 0.78–1.0) using a threshold criterion of 50% of the maximum tidal impedance change.

Conclusion

In the assessment of ventilation inhomogeneity with EIT, functional regions of interest obscure the difference between patients with ARDS and control patients without pulmonary pathologies. The LAEM is preferable to the fROI method when assessing ventilation inhomogeneity.     

Experimental acute Babesia caballi infections: II. Response of platelets and fibrinogen

Ponies were acutely infected with Babesia caballi by inoculation with infected red blood cells (RBCs) containing 1 × 10⁸ and 1 × 10⁹ piroplasms. A series of blood samples taken before and after inoculation were analyzed for platelets and fibrinogen, and the results compared with similar analyses made on challenged, premunized ponies and on equids inoculated with uninfected RBCs. In acutely infected animals there were immediate decreases in platelet counts that persisted at least through Day 18 after inoculation (AI). Concomitantly, plasma fibrinogen levels rose, reaching peak values between Days 6–17. Clot retractions in vitro were impaired in these ponies during Days 9–16. No large diminutions in platelet counts or elevations of fibrinogen levels were observed in the challenged, premunized ponies or the group transfused with uninfected RBCs. In fact, the effect of challenge was to maintain or increase platelet counts. Our results plainly indicate that B. caballi can elicit alterations in clotting factor levels in its hosts during acute infections.